May 20, 2026

Metabolomics for measuring polyamine levels in the plasma, brain and liver of mice

DOI
https://dx.doi.org/10.17504/protocols.io.n2bvj1b1nvk5/v1
  • 1KU Leuven
Hanne HD Dhondt
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DOI: https://dx.doi.org/10.17504/protocols.io.n2bvj1b1nvk5/v1
Protocol Citation: Aisha Sati 2026. Metabolomics for measuring polyamine levels in the plasma, brain and liver of mice . protocols.io https://dx.doi.org/10.17504/protocols.io.n2bvj1b1nvk5/v1
License: This is an open access  protocol  distributed under the terms of the  Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 24, 2026
Last Modified: May 20, 2026
Protocol  Integer ID: 244063
Keywords: ASAPCRN, polyamine levels in the plasma, measuring polyamine level, analysis of polyamine species, metabolomic, polyamine species, using metabolomic, liver sample, liver of mice
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: ASAP-024297
Abstract
This protocol details the analysis of polyamine species in plasma, brain, and liver samples using metabolomics-based approaches.
Materials
Reagents

  • Methanol (80%)
  • Acetonitrile (solvent A)
  • Ammonium formate (10 mM, pH 3.8; solvent B)

Equipment


  • Poroshell 120 HILIC-Z PEEK column (Agilent InfinityLab)
  • Dionex UltiMate 3000 LC System (Thermo Scientific)
  • Q Exactive Orbitrap mass spectrometer (Thermo Scientific)
Methods
Analysis of samples to detect relative values of different polyamine species (ornithine, putrescine, spermidine, spermine, N1-acetylated spermine and N1/8-acetylated spermidine) in the plasma (PND14 and 10 weeks), brain (PND14, 10 weeks and 18 months) and liver (PND14).
Dilute the samples in 80% methanol and run on liquid chromatography-mass spectrometry (LC-MS) (Poroshell 120 HILIC-Z PEEK column (Agilent InfinityLab), Dionex UltiMate 3000 LC System (Thermo Scientific), Q Exactive Orbitrap mass spectrometer (Thermo Scientific)) operated in positive mode.
During LC, carry out a linear gradient with acetonitrile (solvent A) and ammonium formate (10 millimolar (mM) , 3.8 , solvent B) under a constant temperature of 25 °C .

MS operated full scan (range 70-150) with a spray voltage of 3.2 kV, a temperature of 320 °C , sheath gas at 40.00 and auxiliary gas at 10.0.

Set the AGC target at 3x106 using a 70,000 resolution.
Collect the data with Xcalibur software (Thermo Scientific) and analyze by the integration of peak areas.
Acknowledgements
Performed by the VIB metabolomics core at KU Leuven (Dr. B. Ghesquire).